Inhibitors of ADP-ribosylation impair inducible nitric oxide synthase gene transcription through inhibition of NF kappa B activation.
In murine macrophages, inducible NO synthase II (iNOS or NOS-II) is induced at the transcriptional level by IFN-gamma, alone or synergistically with LPS. We investigated the possible role of reactions of ADP-ribosylation in triggering the signaling pathways involved in NOS-II gene expression. Stimulation with IFN-gamma and/or LPS of RAW 264.7 macrophages, transiently transfected with the NOS-II promoter, was inhibited by ADP-ribosylation inhibitors, indicating that they interfered with the signal(s) responsible for NOS-II gene transcription. We therefore explored the effect of these inhibitors on the activity of IRF-1 and NF kappa B transcription factors known to be involved in NOS-II induction by IFN-gamma and LPS. No effect was observed on IRF-1 activation. However, NF kappa B binding to its target sequence diminished and transfection experiments with an NF kappa B-driven reporter plasmid demonstrating that ADP-ribosylation inhibitors suppressed NF kappa B-dependent promoter activity. These results provide evidence that a step involving ADP-ribosylation is required to activate NF kappa B-mediated gene transcription.
Le Page
C
C
Unité 365 INSERM Interférons et Cytokines Institut Curie, Paris, France.
Sanceau
J
J
Drapier
J C
JC
Wietzerbin
J
J
D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors; D004790:Enzyme Induction / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017930:Genes, Reporter / Q000235:genetics; D050856:Interferon Regulatory Factor-1; D007371:Interferon-gamma / Q000037:antagonists & inhibitors / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009536:Niacinamide / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D010750:Phosphoproteins / Q000037:antagonists & inhibitors; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011401:Promoter Regions, Genetic / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection / Q000235:genetics
D001549:Benzamides; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D050856:Interferon Regulatory Factor-1; C494258:Irf1 protein, mouse; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010750:Phosphoproteins; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D014157:Transcription Factors; D009536:Niacinamide; D007371:Interferon-gamma; C025160:3-aminobenzamide; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse